Plant Transformation Via Agrobacterium Tumefaciens

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Plant Transformation via Agrobacterium Tumefaciens

Author : Phetole Mangena
Publisher : CRC Press
ISBN 13 : 1000638154
Total Pages : 244 pages
Book Rating : 4.0/5 (6 download)

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Book Synopsis Plant Transformation via Agrobacterium Tumefaciens by : Phetole Mangena

Download or read book Plant Transformation via Agrobacterium Tumefaciens written by Phetole Mangena and published by CRC Press. This book was released on 2022-09-01 with total page 244 pages. Available in PDF, EPUB and Kindle. Book excerpt: Plant Transformation via Agrobacterium Tumefaciens compiles fundamental and specific information and procedures involving in vitro soybean transformation, which forms the basis for the Agrobacterium-mediated genetic manipulation of soybean using plant tissue culture. This method serves as one of the most preferred, reliable and cost-effective mechanism of transgene expression in both leguminous recalcitrant species and non-legume crops. The technology is favoured due to its simplicity, feasibility and high transformation rates that are so far achieved mostly in monocot plants and a few dicot genotypes. This book provides a comprehensive review of plant transformation which remains necessary for many researchers who are still facing protocol-related hurdles. Among some of the major topics covered in Plant Transformation via Agrobacterium Tumefaciens are the history and discovery of Agrobacterium bacterium, longstanding challenges causing transformation inefficiencies, types and conditions of explants, development of transgenic plants for stress resistance, and the role of transgenic plants on animal/human health, including the environment. Plant Transformation via Agrobacterium Tumefaciens helps the reader to understand how soybean, like many other orphan legume crops, faces the risk of overexploitation which may render the currently available varieties redundant and extinct should its narrow gene pool not improve. Plant transformation serves as a key technique in improving the gene pool, while developing varieties that are drought tolerant, have enhanced nutritional value, pest resistant and reduce the destruction by disease causing microorganims. This book is an essential foundation tool that is available for researchers and students to reinforce the application of Agrobacterium-mediated genetic transformation in soybean.

Plant Tissue Culture Manual

Author : K. Lindsey
Publisher : Springer Science & Business Media
ISBN 13 : 940090181X
Total Pages : 138 pages
Book Rating : 4.4/5 (9 download)

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Book Synopsis Plant Tissue Culture Manual by : K. Lindsey

Download or read book Plant Tissue Culture Manual written by K. Lindsey and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 138 pages. Available in PDF, EPUB and Kindle. Book excerpt: This manual comprises a range of techniques for research workers in the fields of cell and molecular biology, physiology, plant breeding and propagation, and genetic engineering.

Agrobacterium biology and its application to transgenic plant production

Author : Hau-Hsuan Hwang
Publisher : Frontiers Media SA
ISBN 13 : 2889195740
Total Pages : 167 pages
Book Rating : 4.8/5 (891 download)

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Book Synopsis Agrobacterium biology and its application to transgenic plant production by : Hau-Hsuan Hwang

Download or read book Agrobacterium biology and its application to transgenic plant production written by Hau-Hsuan Hwang and published by Frontiers Media SA. This book was released on 2015-06-26 with total page 167 pages. Available in PDF, EPUB and Kindle. Book excerpt: The broad host range pathogenic bacterium Agrobacterium tumefaciens has been widely studied as a model system to understand horizontal gene flow, secretion of effector proteins into host cells, and plant-pathogen interactions. Agrobacterium-mediated plant transformation also is the major method for generating transgenic plants for research and biotechnology purposes. Agrobacterium species have the natural ability to conduct interkingdom genetic transfer from bacteria to eukaryotes, including most plant species, yeast, fungi, and even animal cells. In nature, A. tumefaciens causes crown gall disease resulting from expression in plants of auxin and cytokinin biosynthesis genes encoded by the transferred (T-) DNA. Gene transfer from A. tumefaciens to host cells requires virulence (vir) genes that reside on the resident tumor-inducing (Ti) plasmid. In addition to T-DNA, several Virulence (Vir) effector proteins are also translocated to host cells through a bacterial type IV secretion system. These proteins aid in T-DNA trafficking through the host cell cytoplasm, nuclear targeting, and T-DNA integration. Genes within native T-DNAs can be replaced by any gene of interest, making Agrobacterium species important tools for plant research and genetic engineering. In this research topic, we provided updated information on several important areas of Agrobacterium biology and its use for biotechnology purposes.

Agrobacterium: From Biology to Biotechnology

Author : Tzvi Tzfira
Publisher : Springer Science & Business Media
ISBN 13 : 0387722904
Total Pages : 768 pages
Book Rating : 4.3/5 (877 download)

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Book Synopsis Agrobacterium: From Biology to Biotechnology by : Tzvi Tzfira

Download or read book Agrobacterium: From Biology to Biotechnology written by Tzvi Tzfira and published by Springer Science & Business Media. This book was released on 2007-12-25 with total page 768 pages. Available in PDF, EPUB and Kindle. Book excerpt: Agrobacterium is a plant pathogen which causes the “crown-gall” disease, a neoplastic growth that results from the transfer of a well-defined DNA segment (“transferred DNA”, or “T-DNA”) from the bacterial Ti (tumor-inducing) plasmid to the host cell, its integration into the host genome, and the expression of oncogenes contained on the T-DNA. The molecular machinery, needed for T-DNA generation and transport into the host cell and encoded by a series of chromosomal (chv) and Ti-plasmid virulence (vir) genes, has been the subject of numerous studies over the past several decades. Today, Agrobacterium is the tool of choice for plant genetic engineering with an ever expanding host range that includes many commercially important crops, flowers, and tree species. Furthermore, its recent application for the genetic transformation of non-plant species, from yeast to cultivated mushrooms and even to human cells, promises this bacterium a unique place in the future of biotechnological applications. The book is a comprehensive volume describing Agrobacterium's biology, interactions with host species, and uses for genetic engineering.

Gene Transfer to Plants

Author : Ingo Potrykus
Publisher : Springer Science & Business Media
ISBN 13 : 3642792472
Total Pages : 370 pages
Book Rating : 4.6/5 (427 download)

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Book Synopsis Gene Transfer to Plants by : Ingo Potrykus

Download or read book Gene Transfer to Plants written by Ingo Potrykus and published by Springer Science & Business Media. This book was released on 2013-06-29 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Advances in Wheat Genetics: From Genome to Field

Author : Yasunari Ogihara
Publisher : Springer
ISBN 13 : 4431556753
Total Pages : 421 pages
Book Rating : 4.4/5 (315 download)

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Book Synopsis Advances in Wheat Genetics: From Genome to Field by : Yasunari Ogihara

Download or read book Advances in Wheat Genetics: From Genome to Field written by Yasunari Ogihara and published by Springer. This book was released on 2015-09-15 with total page 421 pages. Available in PDF, EPUB and Kindle. Book excerpt: This proceedings is a collection of 46 selected papers that were presented at the 12th International Wheat Genetics Symposium (IWGS). Since the launch of the wheat genome sequencing project in 2005, the arrival of draft genome sequences has marked a new era in wheat genetics and genomics, catalyzing rapid advancement in the field. This book provides a comprehensive review of the forefront of wheat research, across various important topics such as germplasm and genetic diversity, cytogenetics and allopolyploid evolution, genome sequencing, structural and functional genomics, gene function and molecular biology, biotic stress, abiotic stress, grain quality, and classical and molecular breeding. Following an introduction, 9 parts of the book are dedicated to each of these topics. A final, 11th part entitled “Toward Sustainable Wheat Production” contains 7 excellent papers that were presented in the 12th IWGS Special Session supported by the OECD. With rapid population growth and radical climate changes, the world faces a global food crisis and is in need of another Green Revolution to boost yields of wheat and other widely grown staple crops. Although this book focuses on wheat, many of the newly developed techniques and results presented here can be applied to other plant species with large and complex genomes. As such, this volume is highly recommended for all students and researchers in wheat sciences and related plant sciences and for those who are interested in stable food production and food security.

Plant Genetic Engineering

Author : A.D. Arencibia
Publisher : Elsevier
ISBN 13 : 008053905X
Total Pages : 283 pages
Book Rating : 4.0/5 (85 download)

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Book Synopsis Plant Genetic Engineering by : A.D. Arencibia

Download or read book Plant Genetic Engineering written by A.D. Arencibia and published by Elsevier. This book was released on 2000-02-14 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: Plant biotechnology offers important opportunities for agriculture, horticulture, and the pharmaceutical and food industry by generating transgenic varieties with altered properties. This is likely to change farming practice and reduce the potential negative impact of plant production on the environment. This volume shows the worldwide advances and potential benefits of plant genetic engineering focusing on the third millennium. The authors discuss the production of transgenic plants resistant to biotic and abiotic stress, the improvement of plant qualities, the use of transgenic plants as bioreactors, and the use of plant genomics for genetic improvement and gene cloning. Unique to this book is the integrative point of view taken between plant genetic engineering and socioeconomic and environmental issues. Considerations of regulatory processes to release genetically modified plants, as well as the public acceptance of the transgenic plants are also discussed.This book will be welcomed by biotechnologists, researchers and students alike working in the biological sciences. It should also prove useful to everyone dedicated to the study of the socioeconomic and environmental impact of the new technologies, while providing recent scientific information on the progress and perspectives of the production of genetically modified plants.The work is dedicated to Professor Marc van Montagu.

Plant Transformation Technologies

Author : Charles Neal Stewart
Publisher : John Wiley & Sons
ISBN 13 : 0470958944
Total Pages : 314 pages
Book Rating : 4.4/5 (79 download)

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Book Synopsis Plant Transformation Technologies by : Charles Neal Stewart

Download or read book Plant Transformation Technologies written by Charles Neal Stewart and published by John Wiley & Sons. This book was released on 2011-01-31 with total page 314 pages. Available in PDF, EPUB and Kindle. Book excerpt: Plant Transformation Technologies is a comprehensive, authoritative book focusing on cutting-edge plant biotechnologies, offering in-depth, forward-looking information on methods for controlled and accurate genetic engineering. In response to ever-increasing pressure for precise and efficient integration of transgenes in plants, many new technologies have been developed. With complete coverage of these technologies, Plant Transformation Technologies provides valuable insight on current and future plant transformation technologies. With twenty-five chapters written by international experts on transformation technologies, the book includes new information on Agrobacterium, targeting transgenes into plant genomes, and new vectors and market systems. Including both review chapters and protocols for transformation, Plant Transformation Technologies is vitally important to graduate students, postdoctoral students, and university and industry researchers.

The Tomato Genome

Author : Mathilde Causse
Publisher : Springer
ISBN 13 : 3662533898
Total Pages : 260 pages
Book Rating : 4.6/5 (625 download)

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Book Synopsis The Tomato Genome by : Mathilde Causse

Download or read book The Tomato Genome written by Mathilde Causse and published by Springer. This book was released on 2016-11-21 with total page 260 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book describes the strategy used for sequencing, assembling and annotating the tomato genome and presents the main characteristics of this sequence with a special focus on repeated sequences and the ancestral polyploidy events. It also includes the chloroplast and mitochondrial genomes. Tomato (Solanum lycopersicum) is a major crop plant as well as a model for fruit development, and the availability of the genome sequence has completely changed the paradigm of the species’ genetics and genomics. The book describes the numerous genetic and genomic resources available, the identified genes and quantitative trait locus (QTL) identified, as well as the strong synteny across Solanaceae species. Lastly, it discusses the consequences of the availability of a high-quality genome sequence of the cultivated species for the research community. It is a valuable resource for students and researchers interested in the genetics and genomics of tomato and Solanaceae.

Transgenic Crops of the World

Author : Ian S. Curtis
Publisher : Springer Science & Business Media
ISBN 13 : 1402023332
Total Pages : 448 pages
Book Rating : 4.4/5 (2 download)

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Book Synopsis Transgenic Crops of the World by : Ian S. Curtis

Download or read book Transgenic Crops of the World written by Ian S. Curtis and published by Springer Science & Business Media. This book was released on 2012-08-10 with total page 448 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the first transgenic plants were produced back in the early 1980s, there have been substantial developments towards the genetic engineering of most crops of our world. Initial studies using isolated plant cells and removing their cell walls to form protoplasts, offered the possibility of transferring genetic material by Agrobacterium-mediated gene transfer, chemical agents or electrical charges. However, in those cases were isolated protoplasts could be transformed, often, a shoot regeneration system was not available to induce the production of transgenic plants and any such regenerated plants were subject to mutation or chromosomal of cultured plant organs, such as leaf abnormalities. By the mid-1980s, the use disks, offered the convenience of combining gene transfer, plant regeneration and selection of transformants in a single system. This approach, enabled the production of stable, phenotypically-normal, transgenic potato and tomato plants in culture. By the late 1980s, the use of biolistics offered a means of inserting foreign genes into plant cells which where inaccessible to Agrobacterium infection. Even today, this technology is now standard practice for the production of some transgenic plants.

The Plant Cytoskeleton

Author : Bo Liu
Publisher : Springer Science & Business Media
ISBN 13 : 1441909877
Total Pages : 333 pages
Book Rating : 4.4/5 (419 download)

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Book Synopsis The Plant Cytoskeleton by : Bo Liu

Download or read book The Plant Cytoskeleton written by Bo Liu and published by Springer Science & Business Media. This book was released on 2010-11-23 with total page 333 pages. Available in PDF, EPUB and Kindle. Book excerpt: Plant cells house highly dynamic cytoskeletal networks of microtubules and actin microfilaments. They constantly undergo remodeling to fulfill their roles in supporting cell division, enlargement, and differentiation. Following early studies on structural aspects of the networks, recent breakthroughs have connected them with more and more intracellular events essential for plant growth and development. Advanced technologies in cell biology (live-cell imaging in particular), molecular genetics, genomics, and proteomics have revolutionized this field of study. Stories summarized in this book may inspire enthusiastic scientists to pursue new directions toward understanding functions of the plant cytoskeleton. The Plant Cytoskeleton is divided into three sections: 1) Molecular Basis of the Plant Cytoskeleton; 2) Cytoskeletal Reorganization in Plant Cell Division; and 3) The Cytoskeleton in Plant Growth and Development. This book is aimed at serving as a resource for anyone who wishes to learn about the plant cytoskeleton beyond ordinary textbooks.

Genetic Engineering of Plants

Author : National Research Council
Publisher : National Academies Press
ISBN 13 : 0309034345
Total Pages : 97 pages
Book Rating : 4.3/5 (9 download)

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Book Synopsis Genetic Engineering of Plants by : National Research Council

Download or read book Genetic Engineering of Plants written by National Research Council and published by National Academies Press. This book was released on 1984-02-01 with total page 97 pages. Available in PDF, EPUB and Kindle. Book excerpt: "The book...is, in fact, a short text on the many practical problems...associated with translating the explosion in basic biotechnological research into the next Green Revolution," explains Economic Botany. The book is "a concise and accurate narrative, that also manages to be interesting and personal...a splendid little book." Biotechnology states, "Because of the clarity with which it is written, this thin volume makes a major contribution to improving public understanding of genetic engineering's potential for enlarging the world's food supply...and can be profitably read by practically anyone interested in application of molecular biology to improvement of productivity in agriculture."

Biolistic DNA Delivery in Plants

Author : Sachin Rustgi
Publisher : Humana
ISBN 13 : 9781071603581
Total Pages : 334 pages
Book Rating : 4.6/5 (35 download)

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Book Synopsis Biolistic DNA Delivery in Plants by : Sachin Rustgi

Download or read book Biolistic DNA Delivery in Plants written by Sachin Rustgi and published by Humana. This book was released on 2021-04-25 with total page 334 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume details protocols for the use of the biolistic DNA delivery method in different plant species. Chapters guide readers through non-protocol chapters that cover relevant topics of interest, a broad overview of the field, exciting modifications of the system, and reliable plant transformation procedures in different plant species. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Biolistic DNA Delivery: Methods and Protocols aims to provide a comprehensive collection of protocols to intended to be a practical guide for the novice as well as the advanced user in the field of plant genetic transformation.

Methods In Arabidopsis Research

Author : Nam-hai Chua
Publisher : World Scientific Publishing Company
ISBN 13 : 9813103426
Total Pages : 496 pages
Book Rating : 4.8/5 (131 download)

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Book Synopsis Methods In Arabidopsis Research by : Nam-hai Chua

Download or read book Methods In Arabidopsis Research written by Nam-hai Chua and published by World Scientific Publishing Company. This book was released on 1992-06-12 with total page 496 pages. Available in PDF, EPUB and Kindle. Book excerpt: One of the ways to make consistent progress in a particular field of biology consists in choosing a good model system on which to focus the experimental efforts of the scientific community. It has taken a long time for scientists interested in various aspects of the life of plants to reach some sort of consensus. With the advent and impact of molecular biology, the small weed Arabidopsis is now the object of rapidly growing scientific attention. Since it is reasonable to assume that the general molecular mechanisms that are responsible for the physiological, cellular and biochemical properties of plants will be essentially conserved in all plants, it follows that these mechanisms should also operate in Arabidopsis and hence that its genome should contain most of the genes that we need to know about if we want to understand the genetic determination of the life processes in plants.Arabidopsis has a small genome and well documented genetic studies are available. It is easy to grow in large numbers and mutants defining important genetically controlled mechanisms are either available, or can readily be obtained. Various methods to introduce and express isolated homologous or heterologous genes are available. It is therefore realistic and desirable to aim at exploring the genome of this plant in very great detail. As will be illustrated in this book all the elements for such a grand strategy are in place.More and more scientists are therefore willing to accept the obvious and very real practical disadvantages resulting from its small size when experiments call for the isolation of proteins, membranes, subcellular fractions etc, in order to benefit from its extraordinary experimental advantages as a model system in molecular genetics. One can safely predict that in the next decade studies with Arabidopsis will provide major breakthroughs in our understanding of most aspects of plant physiology and developmental biology. The importance of this knowledge for plant breeding and therefore for a sustainable highly productive agriculture cannot be overestimated. We therefore expect that this book will provide valuable guidelines to all those who are planning experiments aimed at understanding various aspects of plant growth, productivity and interactions with the environment. The book offers a wealth of methodical and theoretical information as well as valuable references. It should be of use to students, teachers, as well as advanced researchers and those breeders who want to use molecular techniques in breeding.

Agrobacterium Protocols

Author : Kan Wang
Publisher : Springer Science & Business Media
ISBN 13 : 1597451312
Total Pages : 473 pages
Book Rating : 4.5/5 (974 download)

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Book Synopsis Agrobacterium Protocols by : Kan Wang

Download or read book Agrobacterium Protocols written by Kan Wang and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 473 pages. Available in PDF, EPUB and Kindle. Book excerpt: Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agric- tural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Ag- bacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent devel- ments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.

Agrobacterium Tumefaciens

Author : Eugene W. Nester
Publisher : American Phytopathological Society
ISBN 13 :
Total Pages : 344 pages
Book Rating : 4.F/5 ( download)

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Book Synopsis Agrobacterium Tumefaciens by : Eugene W. Nester

Download or read book Agrobacterium Tumefaciens written by Eugene W. Nester and published by American Phytopathological Society. This book was released on 2005 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: This anthology traces the fascinating progress from plant pathology to biotechnology through 38 scientific papers on Agrobacterium, published over the past century. Included are the seminal scientific papers on the biology and application of Agrobacterium with introductory commentaries mostly by those involved in the original work. The commentaries give background to the papers and explain the problems faced and the techniques used, providing insight into the way fundamental research progresses.Agrobacterium tumefaciens has played a major role in the astounding advances that have been made over the past several decades in the areas of plant genetics, plant molecular biology, and plant genetic engineering. The papers included in this book were integral to the current understanding of the interaction of Agrobacterium with its hosts, its development into a major player in the genetic engineering of plants, and the biological control of crown gall.Agrobacterium tumefaciens: From Plant Pathology to Biotechnology is divided into five sections. The first section begins with 1904 when Erwin F. Smith began detailed work on crown gall and considered it to be a plant pathological problem. It explores many of the biological discoveries made over the past century, including the pivotal moment when Armin C. Braun discovered that crown gall was a plant cancer. Other papers cover the beginnings of T-DNA research and the development of vectors to improve the process of transferring T-DNA from bacterium to plant cell. The second section delves further into vector systems and genetic coding for disease and insect resistance, exploring the evolution of genetic engineering in crops.The final three sections deal with themes developed from crown gall studies, including ?quorum sensing? or population density, the DNA sequencing of one strain of A. tumefaciens, and the first genetically engineered organism, strain K1026, released for commercial use.According to Editor, Eugene Nester, ?This book should serve as a testimony to the 100 years of research on this remarkable organism, as well as to an international group of investigators who helped reveal secrets of this natural genetic engineer.? Students, professors, plant pathologists, microbiologists, or anyone interested in research and/or the history of plant pathology and biotechnology, will find this collection of papers an intriguing read.From the Preface: ??the journey is not over. As the commentary by Paul Hooykaas indicates, it looks as though T-DNA will insert into any cell, be it plant, fungal or even mammalian. Is there a possibility of using Agro?bacterium in gene therapy? Will Agrobacterium prove to be as useful a tool in fungal genetics as it has been in plant genetics? Its potential is mind-boggling.?

Transgenic Wheat, Barley and Oats

Author : Huw D. Jones
Publisher : Humana Press
ISBN 13 : 9781588299611
Total Pages : 0 pages
Book Rating : 4.2/5 (996 download)

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Book Synopsis Transgenic Wheat, Barley and Oats by : Huw D. Jones

Download or read book Transgenic Wheat, Barley and Oats written by Huw D. Jones and published by Humana Press. This book was released on 2008-11-11 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Understanding the physical and genetic structure of cereal genomes and how defined coding and non-coding regions interact with the environment to determine a phenotype are key to the future of plant breeding and agriculture. The production and characteri- tion of transgenic plants is a powerful reverse genetic strategy increasingly used in cereals research to ascribe function to defined DNA sequences. However, the techniques and resources required to conduct these investigations have, until recently, been difficult to achieve or totally lacking in wheat, barley and oat. This book brings together the l- est protocols for the transformation, regeneration and selection using both biolistic and Agrobacterium tumefaciens appropriate for these three species. It includes two chapters describing in vitro Agrobacterium co-cultivation, one leading to germ line transformation with no need for tissue culture-based regeneration. In addition, it has several chapters dedicated to the manipulation of gene expression and characterisation of the recombinant locus and transgenic plants. Finally, it tackles the issues of GM risk assessment, field trials and substantial equivalence in terms of transcriptomics, proteomics and metabolomics. Although this book is dedicated to the temperate small grain cereals wheat, barley and oats, many of the techniques described could be readily adapted for other cereals or plants generally. We thank all the contributing authors for their timely and informative chapters, the staff of Humana Press, especially John Walker for their guidance, and Helen Jenkins for her proof-reading, word processing and administrative support. v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix PART I.